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To prepare primary neutrophil CM, 12 10 6 freshly procured neutrophils (LDN and HDN) were washed with HBSS and cultured overnight in serum-free RPMI or AIM-V medium in the presence or absence of 1 M nicotine

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But its also possible that it has an effect over time

The range is broad because the level of nicotine depends on many different factors, such as how the tobacco is grown and processed, the size of the cut of the leaves, and the pH of the product